An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues.
Formalin-fixed paraffin-embedded tissues (FFPET) represent the largest source of archival biological material available for genomic studies. In this work we present an advanced protocol for extraction of high quality DNA from FFPET that can be applied in several molecular studies. Although cat mammary tumours (CMT) are the third most frequent tumour in cats the recovery of significant number of samples for molecular studies are in some way restricted to FFPET samples. We were able to obtain high quality DNA from FFPET of thirty six CMT that were subjected to pre-fixation and fixation processes routinely used in the veterinary hospitals. The quality of DNA obtained was tested by PCR amplification using six sets of primers that amplify single-copy fragments. The DNA fragments obtained were further sequenced. This protocol was able to provide FFPET gDNA that can be amplified and sequenced for larger fragments up to 1182bp.
Santos
Sara
S
Institute for Biotechnology and Bioengineering, Centre of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (CGB-UTAD/IBB), Quinta de Prados, Apdo 1013, 5001-801 Vila Real, Portugal.
Sá
Daniela
D
Bastos
Estela
E
Guedes-Pinto
Henrique
H
Gut
Ivo
I
Gärtner
Fátima
F
Chaves
Raquel
R
D000818:Animals; D002371:Cat Diseases / Q000473:pathology; D002415:Cats; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004273:DNA, Neoplasm / Q000235:genetics* / Q000302:isolation & purification*; D005260:Female; D005557:Formaldehyde; D016678:Genome; D015674:Mammary Neoplasms, Animal / Q000473:pathology*; D008970:Molecular Weight; D021141:Nucleic Acid Amplification Techniques; D016612:Paraffin Embedding
D004273:DNA, Neoplasm; D005557:Formaldehyde; D004247:DNA